Dissertation / PhD Thesis FZJ-2014-00850

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png
Interdomain Functional Dynamics of Phosphoglycerate Kinase Studied by Single-Molecule FRET



2014
Forschungszentrum Jülich GmbH Zentralbibliothek, Verlag Jülich
ISBN: 978-3-89336-943-0

Jülich : Forschungszentrum Jülich GmbH Zentralbibliothek, Verlag, Schriften des Forschungszentrums Jülich. Reihe Schlüsseltechnologien / Key Technologies 81, V, 179 S. () = Heinrich-Heine-Universität Düsseldorf, Diss., 2013

Please use a persistent id in citations:

Abstract: The thesis reflects the steps followed to perform reliable single-molecule FRET (smFRET) experiments on phosphoglycerate kinase (PGK) from yeast. The main text is essential and divided in short chapters in order to relieve the reading. All technical details, demostrations, long descriptions, protocols, figures, and tables are reported in the appendix for the reader more interested in a deeper understanding. The first part encompasses Chap.3 to Chap.9 and is dedicated to the establishment of reliable single-molecule fluorescence measurements. Here, it is condensed the Ph.D. experience in a manual-like form for newcomers of the fieldwilling to perform single-molecule experiments from scratch. Since high quality and controlled labeled samples are a fundamental prerequisite for single-molecule fluorescence measurements, this topic is discussed in Chap.3. The theoretical background for the comprehension of singlemolecule detection in solution is given in a compact and consistent form in Chap.4. The basics about the three principal experimental approaches needed to perform single-molecule fluorescence measurements are presented in Chap.5, Chap.6 and Chap.7. The data treatment of the smFRET experiments is explained in Chap.8 and is mainly based on the works ofGopich and Szabo, and Seidel and co-workers. The procedure applied to determine the set of parameteres required for the analysis of the data is found in Chap.9. Here, a new strategy for the measurement of precise relative quantum yields on the confocal microscope is reported. Finally, in the last chapter (Chap.10) the whole knowledge is applied to study the interdomain dynamics of PGK. Here, the protein motions are modeled with an elastic network simulated under a random Multiparticle Collision Dynamics (rMPC) approach.

Keyword(s): Dissertation


Note: Heinrich-Heine-Universität Düsseldorf, Diss., 2013

Contributing Institute(s):
  1. Molekulare Biophysik (ICS-5)
Research Program(s):
  1. 452 - Structural Biology (POF2-452) (POF2-452)

Appears in the scientific report 2013
Database coverage:
OpenAccess
Click to display QR Code for this record

The record appears in these collections:
Institute Collections > IBI > IBI-6
Document types > Theses > Ph.D. Theses
Workflow collections > Public records
ICS > ICS-5
Publications database
Open Access

 Record created 2014-01-27, last modified 2021-03-08


OpenAccess:
Download fulltext PDF
External link:
Download fulltextFulltext by OpenAccess repository
Rate this document:

Rate this document:
1
2
3
 
(Not yet reviewed)